In order to investigate the mutation of HBV polymerase gene reverse transcription conserved region (P region) in chronic hepatitis B (CHB) patients, 212 CHB patients who took antiretroviral treatment with nucleotide analogues were chosen. The drug resistance mutations of HBV P region and HBV genotype were detected by Pyrosequencing. Sequence analysis showed that the drug resistance sites of HBV P region located at sites 173; 180; 181; 184; 204; 236 and 250. The main site of HBV P region drug resistance was 204 and 180, accounting for 35.8% and 23.5%, respectively. There were significant differences in the mutation rate of site 180 among different age groups. There were also significant differences in the mutation rate of site 204 among younger than 30 age group, 41 to 50 age group and 51 to 60 age group. (P < 0.05, P < 0.01). The mutation rate of site 180 combined with site 204 was 66.6%. The mutation rate of site 181 combined with site 236 was 23.3%. The age of C genotype infected patients was significantly older than B genotype infected patients (P < 0.01). M204V/I mutation mostly existed in the form of joint L180M mutation, the mutation rate was age-related. The detection of HBV genotypes and drug resistance sites of HBV P region have important clinical implications for the treatment and prognosis of patients with CHB.
Adult ; Aged ; Antiviral Agents ; pharmacology ; China ; Drug Resistance, Viral ; Female ; Gene Products, pol ; genetics ; Genotype ; Hepatitis B virus ; classification ; drug effects ; enzymology ; genetics ; Hepatitis B, Chronic ; drug therapy ; virology ; Humans ; Male ; Middle Aged ; Mutation, Missense ; Young Adult

Objective: To learn the genotypic drug resistance of men who have sex with men ( MSM ) with HIV who failed in antiviral therapy in Yunnan Province, in order to provide basis for improving the effect of antiviral therapy. Methods: The patients who were infected with HIV-1, homosexual transmitted and failed in antiviral therapy in Yunnan Province from 2014 to 2019 were recruited. Their plasma samples were tested by reverse transcription nested polymerase chain reaction ( RT-nPCR ) , the fragments were spliced using ContigExpress, and the resistance to 8 protease inhibitors ( PIs ) , 7 nucleoside reverse transcriptase inhibitors ( NRTIs ) and 5 non-nucleoside reverse transcriptase inhibitors ( NNRTIs ) were obtained from the HIV drug resistance data website of Stanford University. Results: A total of 205 HIV/AIDS cases were included, 169 positive plasma samples were amplified, 112 cases were drug resistant, and the rate of drug resistance was 66.27%. The patients who were aged 30-49 years ( 76.09% ) , had genotype of CRF01_AE ( 76.34% ) or treated by AZT+3TC+NVP ( 77.08% ) had higher resistance rate. The resistance rates of NNRTIs, NRTIs and PIs were 62.72%, 49.70% and 2.96%, respectively; the resistance rates of NVP and EFV in NNRTIs were 62.72% and 61.54%. The main mutation site associated with NNRTIs was K103, accounting for 21.89% ( 37 cases ) ; the main mutation site associated with NRTIs was M184, accounting for 39.64% ( 67 cases ) ; the main mutation sites associated with PIs were M46L/K, accounting for 2.96% ( 5 cases ) , resulting in high resistance to NFV. Conclusions The drug resistance rate of HIV-infected MSM with failure of antiviral therapy in Yunnan Province is relatively high, with CRF01_AE as the main gene subtype of drug resistance. The drug resistance rate of NNRTIs is relatively high, especially NVP and EFV.

Objective To evaluate the long-term efficacy of antiretroviral therapy and drug resistance among human immunodeficiency virus-1(HIV-1)-infected children in Yunnan.Methods In this retrospective study,CD4+T cell counts,HIV viral loads and genetic drug resistance results were obtained from HIV-1-infected children who were treated with antiretroviral treatment between January 2004 to July 2015.Results A total of 1 078 HIV/acquired immune deficiency syndrome(AIDS)children were treated with antiviral therapy.Before treatment,the average CD4+cell number was(466.8 ± 397.2)cells/μL. The percentages of children with CD4+cell count >750 cells/μL after 1-year,3-year,5-year and 8-year treatment were 54.31%,62.87%,68.46% and 74.19%,respectively.Virological failure occurred in 150 HIV/AIDS children(13.9%),and the virological failure rate was 4.3/100 child-years.Among those 150 patients with virological failure,104 cases(69.33%)exhibited genetic resistance to antiretroviral drugs.The prevalent mutations associated with drug resistance were M 184V/I(75.0% [78/104]), K103N(43.3%[45/104]),G190A(29.8%[31/104]),Y181C(22.1%[23/104]),T215Y/F(20.2%[21/104]).Conclusions After long-term antiretroviral treatment,most of the HIV-infected children have restored the immunity and suppressed HIV viral replication successfully.HIV resistance is the main cause of virological failure.Drug resistance mutations mainly occur in nucleoside reverse transcriptase inhibitor and non-nucleoside reverse transcriptase inhibitor,and the resistance rate of proteinase inhibitor is low.Early genetic resistance testing and switch to second-line therapy will improve the treatment outcome.

Objective To understand the HIV infection status in middle aged and old men and the association between HIV infection and female sex workers in a city in Hubei province and provide evidence for the targeted prevention and control of HIV infection.Methods Ten communities/villages were selected through stratified cluster sampling from the suburban and rural areas of the city.A total of 634 men aged ≥50 years were surveyed by using self-designed questionnaire in December,2014.Results The awareness rate of HIV/AIDS related knowledge was 13.3％ (68/513).Among the men surveyed,63.2％ (371/587) thought they couldn't be infected with HIV,9.1％ (52/573) believed that extramarital sex is acceptable,and 32.4％ (189/583) considered that commercial sexual behavior is just as well.Among the surveyed men reporting sexual behavior in the past 6 months,6.6％ (15/228) had sex with regular sexual partners,in which 63.6％ (7/11) never used condoms,21.8％(128/586) reported that some friends had commercial sexual behaviors.Among 601 men receiving HIV testing,7 were HIV positive (1.2％).The HIV detection rates among the men living at 3 communities near red light district were 4.2％ (5/119),1.1％ (1/95) and 0.5％ (1/192) respectively;while no HIV infections were found in the men living at 2 towns far away from red light district.Seventy seven of 581 men surveyed (13.2％) reported to meet visiting female sex workers and they had higher HIV positive rate (5.2％,4/77) than those without such experiences (0.6％,3/504) (P=0.007).Conclusion The middle aged and old men in this city had low awareness of HIV/AIDs related knowledge,and high-risk sexual behaviors existed among them.HIV positive rate was relatively high in this population,and HIV infection might be associated with sex service.

Objective To investigate the impact flat deformity of the cerebral cortex induced by congenital hydrocephalus on motor functioning and cognition.Methods Tomography was used to confirm the presence of flat cortex deformity in three congenital hydrocephalus patients ranging in age from 20 to 33.Their motor control,balance,cognition and ability in the activities of daily living (ADL) were evaluated using the Fugl-Meyer assessment (FMA),the Berg balance scale (BBS),the mini-mental state examination (MMSE) and the modified Barthel index (MBI).Speech dysfunction was judged on the basis of clinical communication.The patients were scanned using a 64-slice spiral CT and size-of-ventricle indices were calculated.Results All 3 patients underwent ventrideperitoneal shunt.Their FMA scores were 75.5,83 and 100,with BBS scores of 4,24 and 56,MMSE scores of 14,23 and 26,and the MBIs of 40,90 and 100.CT images showed obvious ventricle enlargement and a thinner cortex layer in all three,with the thinnest part 0.18 cm,0.22 cm and 0.57 cm.Their ventricle indexes were 303％,288％ and 192％,respectively.Conclusion Although there is no systematic rehabilitation therapy for such patients,their motor,cognition and speech functioning were good,indicating the great potential for plasticity of the human brain.

Objective To investigate the subtypes distribution of human immunodeficiency virus (HIV)-1 epidemic strains and the characteristics of amino acid variation in different areas of Yunnan Province.Methods Totally 800 HIV/AIDS plasma specimens and epidemiological information were collected between 2012 and 2015 from 14 areas of Yunnan.Viral RNA was extracted and amplified using RT polymerase chain reaction (PCR).4.5 kb 5'halves fragments were obtained and directly sequenced.Subtypes of strains were identified by Genotyping,MEGA 6.06 and BLAST.Grouping was analyzed by location and subtype.Entropy software was used to analyze the difference of amino acid sequences between different groups according to the sampling location and subtypes to analyze the regional distribution and genetic variation of HIV-1 subtypes in Yunnan Province.Results Of the 800 plasma specimens,a total of 446 genomic sequences from 12 areas were successfully amplified and sequenced.After genotypes were identified,the subtypes of HIV-1 strains prevalent in Yunnan were CRF08_BC (58.3％),CRF01_AE (19.3％),CRF07_BC (11.6％),unknown recombinant forms (7.1％),B(B') (1.7％) and C (1.3％).The geographical distribution in Yunnan was analyzed.The CRF01_AE predominated in Dehong,Xishuangbarma and Wenshan.The CRF08_BC predominated in Lincang,Honghe and Puer (more than 70.0％) and CRF08_BC was prevalent in the other areas.But CRF07_BC in Kunming,Yuxi and Dali accounted for more than 20 ％.The constitutions of amino acid of three majors CRF08_BC,CRF01 _AE and CRF07_BC were different on 17,14 and 18 amino acid sites with statistical differences in the eastern and western regions of Yunnan Province (均P ＜ 0.05).Conclusions HIV-1 strains transmit and vary genetically in the province widely.The amino acid mutation sites of eastern and western strains are different.This difference represents that the same subtype strains in different geographical distribution vary on different genetic background and are selected by immune responses.The epidemic trends need to be closely monitored.

Objective To analyze the genetic structure and recombination characteristics of a new-ly discovered HIV-1 unique recombinant strain in Yunnan Province. Methods During a test for drug-resist-ant HIV genotypes in Yunnan Province in 2016, a recombinant fragment was found in the pol region of a HIV-1 strain isolated from a patient. Two overlapping segments of the HIV-1 genome were amplified by RT-PCR, and then the products were sequenced. Recombination analysis was performed using RIP, jpHMM and SimPlot3. 5 software. A phylogenetic tree was constructed for homology analysis by Neighbor-joining method using MEGA6. 06 software. Results A nearly full-length HIV-1 gene sequence with 8590 bp in length was obtained. Breakpoint analysis indicated that the sequence consisted of CRF01_AE and fragments of B and C subtypes. CRF01_AE was used as the backbone with B and C subtype fragments inserted. The positions were 791 to 1171 for CRF01_AE, 1172 to 2652 for C subtype fragment, 2653 to 2977 for B subtype frag-ment, and 2978 to 9380 for CRF01_AE using HIV-1 HXB2 as the reference strain. Conclusions Some new strains formed by cross-recombination of CRF01_AE and B and C subtypes were discovered in Yunnan Province in recent years. It was found that the recombination pattern of the newly discovered strain was com-plex, suggesting that close attention should be paid to the changes in epidemic trends, which was of great im-portance to understand the current prevalence and epidemic trends of HIV-1.

Objective To investigate the influence factors of mortality among human immunodeficiency virus (HIV)-infected children under highly active antiretroviral therapy (HAART).Methods Retrospective cohort study of 652 children initiated HAART from 2005 to 2014 was conducted,and enrolled patients were followed-up until December,2015.Survival data was analyzed using Kaplan-Meier method and Cox regression model was used to identify independent predictors of mortality among these children on HARRT.Chi-square test and Fisher's exact test were used for comparison between groups.Results Overall,26 of the children died over a follow-up period of 3 116.24 child-years,with a mortality rate of 0.83 per 100 child-years.Twelve (46％)of deaths occurred during the first six months after starting HAART.Cox regression analysis of variables showed that the World Health Organization (WHO) clinical stages Ⅲ/Ⅳ (hazard rate [HR] =10.717,95％confidence interal [95％ CI]:4.189-4.749,P =0.000),baseline hemoglobin ＜ 80 g/L (HR =14.768,95 ％ CI:5.721-38.125,P =0.000),tuberculosis co-infection (HR =4.794,95％ CI:2.105-10.918,P =0.000),baseline CD4+T lymphocyte ＜ 50 cells/μL (HR =4.219,95％ CI:1.524-11.680,P =0.006),weight-for-age z-score ＜-2 (HR =2.983,95 ％ CI:1.094-8.135,P =0.033) were independently associated with death,whereas the age ＜ 7 year-old at HAART initiation was protectire (HR =0.293,95％ CI:0.126-0.684,P =0.005).Conclusions The mortality of children receiving HAART is strongly associated with WHO stages Ⅲ/Ⅳ,hemoglobin ＜ 80 g/L,weight-for-age z-score ＜-2,tuberculosis co-infection and older age at treatment.

Objective:To analyze the variation characteristics of gag gene sequence of human immunodeficiency virus type 1 (HIV-1) epidemic strains in four major acquired immunodeficiency syndrome (AIDS) endemic areas in Yunnan Province. Methods:A total of 480 human immunodeficiency virus (HIV)/AIDS patients from designated antiviral treatment institutions in Kunming City, Dehong Dai and Jingpo Autonomous Prefecture, Hani-Yi Autonomous Prefecture of Honghe and Lincang City in Yunnan Province from January 2019 to December 2020 were randomly selected. The epidemiological information of the patients was collected. The plasma samples were collected and sent to the Yunnan Provincial Infectious Diseases Hospital for analysis. HIV-1 gag gene was amplified by nested polymerase chain reaction for genotyping.The phylogenetic tree was constructed by MEGA 6.06, and the characteristic amino acids were analyzed by VESPA online analysis tool. The gene distances of gag and matrix protein p17, capsid protein p24, nucleocapsid protein p7 and p6 protein of gag protein were calculated by Distance program. The ratio of synonymous mutation frequnency and non-synonymous mutation frequnency (Ks/Ka) was analyzed by SNAP program. Statistical comparison among multiple groups was performed using analysis of variance. Results:The gag gene sequences were successfully obtained from 404 patients.Most of these patients were men (250 cases, 61.9%), 59.7%(241/404) of patients were aged 40 to 59 years, and the main transmission route was heterosexual transmission (61.4%, 248/404). The main epidemic subtypes of HIV-1 were circulating recombinant form(CRF)08_BC (38.4%, 155/404), CRF01_AE (18.3%, 74/404), unique recombinant form (URF) (12.9%, 52/404), CRF07_BC(9.7%, 39/404), C subtype (8.4%, 34/404), other subtypes (6.9%, 28/404) and B subtype (5.4%, 22/404). Two main spreading clusters were formed by CRF08_BC in the phylogenetic tree, and there were significant differences in the distribution of characteristic amino acids of eight loci between the two spreading clusters, including No.79, 93, 121, 122, 151, 363, 395 and 396. The gag gene distances of CRF01_ AE, CRF07_ BC and CRF08_ BC were 0.090±0.004, 0.088±0.004 and 0.078±0.002, respectively, and the difference was statistically significant ( F=118.33, P<0.001). The Ks/Ka values of gag of the three subtypes were 4.003±1.309, 4.141±0.860 and 4.514±1.215, respectively, and the difference was statistically significant ( F=1.35, P<0.001). The Ks/Ka values of p17 were 2.590±0.186, 2.831±0.496 and 2.936±0.475, respectively, and the difference was statistically significant ( F=1.59, P<0.001). The Ks/Ka values of p24 were 12.579±1.116, 10.185±0.494 and 8.522±0.595, respectively, and the difference was statistically significant ( F=1.61, P<0.001). The Ks/Ka values of p7 were 10.850±0.711, 9.717±0.932 and 8.522±0.026, respectively, and the difference was statistically significant ( F=4.24, P<0.001). The Ks/Ka values of p6 were 3.122±0.134, 3.040±1.498 and 4.841±0.353, respectively, and the difference was statistically significant ( F=10.68, P<0.001). Conclusions:CRF08_ BC is the main subtype in these four areas in Yunnan Province.The different clusters of CRF08_BC result in a different pattern of amino acid composition.The variation degree of different subtype gag gene is different in each section. Surveillance of HIV variant strains should be strengthened to control the epidemics of HIV.

Objective:To investigate the effect of sequential suture and adhesion on craniomaxillofacial skin contusion and laceration.Methods:A total of 189 patients with craniomaxillofacial skin contusion and laceration (CMFSCL) were randomly divided into three groups: 66 cases in SSA group, 63 cases in CS group and 60 cases in TS group. Operation time, visual analogue scale (VAS), Vancouver scar scale (VSS) and adverse reactions incidence were compared and analyzed between the three groups. Effect and satisfactory scale were evaluated.Results:Operation time in SSA group (10.67±1.26) min was significantly less than that in CS (18.91±1.38) min and TS group (17.96±1.43) min ( P<0.05). VAS in SSA group 24 h post-operation (3.11±1.01) was significantly lower than that in CS and TS group ( P<0.05). VSS in SSA group 6 months post-operation (1.18±0.21) was significantly lower than that in CS (3.78±1.01) ( P<0.05) and TS group (5.98±1.06) ( P<0.01). Total effective rate of SSA group (96.5%) was significantly higher than that in CS (85.7%) ( P<0.05) and TS group (56.1%) ( P<0.01); total effective rate in CS group was significantly higher than that in TS group ( P<0.05). Infection and dehiscence rates in SSA group were lower than those in CS and TS group ( P<0.01). Satisfactory rate of SSA group (99%) was significantly higher than that of CS (89.1%) and TS group (71.3%) ( P<0.05); the satisfactory rate of CS group was significantly higher than that of TS group ( P<0.05). Conclusions:Sequential suture and adhesion technique is simple and effective for craniomaxillofacial skin contusion and laceration, which is worthy of clinical promotion.